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Bode Ireti Shobayo,1,2,&ast; Victor Saah Taylor,1,&ast; Bluefin Masell Freeman,1,2 James Saysay Kokro,2 Sampson K P Chea1 1Department of Biological Sciences, University of Liberia, Louisiana, Liberia; 2Division of Public Health and Medical Research, National Public Health Institute of Liberia, Monrovia, Liberia&ast;These authors contributed equally to this workCorrespondence: Bode Ireti Shobayo, Division of Public Health and Medical Research, National Public Health Institute of Liberia, Monrovia, Liberia, Tel +231775912252, Email bodeishobayo@gmail.com; shobayobi@ul.edu.lrBackground: Globally, healthcare systems are currently facing a significant challenge in terms of antibiotic resistance. Healthcare professionals actively participate in the process of prescribing, dispensing and administering antibiotics.Objective: We examined the knowledge, attitudes and practices regarding antibiotic use and antibiotic resistance among healthcare professionals working in a tertiary hospital located in Monrovia, Liberia.Methods: A hospital-based cross-sectional survey was carried out from January to June, 2023 involving 61 healthcare workers at the ELWA Hospital, Liberia. A purposive sample of healthcare workers across diverse professional roles was surveyed using a structured questionnaire on antibiotic use and resistance. Data were analyzed in SPSS v25 using descriptive statistics to summarize participant characteristics and inferential tests to explore variable associations.Results: Participants ages ranged from 20 to 60 years (mean = 40.7 ± 5) and nurses constituted the majority professional group (59%). Most respondents (68.9%) disagreed that antibiotics are effective against viral infections (OR = 0.45; p = 0.020). However, 36.1% believed antibiotics could be stopped when symptoms resolve and 24.6% believed leftover antibiotics could be reused (OR = 0.33; p = 0.002). Majority, 72.1% and 70.5%, reported never using antibiotics for body pain or headaches, respectively (OR = 3.67; p = 0.001 and OR = 4.78; p < 0.001). Despite this, 39.3% admitted to sometimes or always storing leftover antibiotics and 39.3% agreed or strongly agreed that stopping antibiotics early is safe (OR = 0.36; p = 0.016).Conclusion: The study identified persistent gaps in healthcare workers’ knowledge, attitudes, and practices regarding antibiotic use and resistance, despite encouraging awareness in some areas. Misconceptions such as premature discontinuation and reuse of leftover antibiotics were common. Findings underscore the need for targeted education and strengthened stewardship programs in Liberia’s healthcare settings.Plain Language Summary: This study looked at how healthcare workers in Monrovia, Liberia understand and use antibiotics. It focused on their knowledge, beliefs, and everyday practices related to antibiotic use and resistance, a growing global health problem. Researchers surveyed 61 healthcare workers at ELWA Hospital, a major health facility in Monrovia. Participants included nurses, doctors, pharmacists, lab technicians, and midwives. They answered a structured questionnaire designed to measure what they know, how they feel, and what they do when it comes to antibiotics.The study found that most healthcare workers knew that antibiotics do not work against viruses and should not be used for things like headaches or body pain. However, many still held incorrect beliefs like thinking it is okay to stop antibiotics early or reuse leftover medicine. Nearly 40% admitted to storing leftover antibiotics, and some disposed of them improperly. On the positive side, most participants followed infection prevention guidelines and recognized antibiotic resistance as a serious public health threat.Antibiotic resistance makes infections harder to treat and is especially dangerous in places with limited healthcare resources. Healthcare workers play a key role in preventing resistance, so it is important they have accurate information and follow best practices. The study shows that while awareness is improving, there are still gaps that need to be addressed. Targeted training and stronger hospital policies can help healthcare workers use antibiotics more responsibly and protect public health in Liberia.Keywords: knowledge, attitudes, practices, antibiotic resistance, antibiotic use, healthcare workers, Liberia